[Vector-mediated shRNA inhibits HIF-1alpha expression in prostate cancer cells]

Hao Ping, Nian-Zeng Xing, Xiao-Chun Chen, Jun-Hui Zhang, Yan-Yong, Yong Yang
Zhonghua Nan Ke Xue, National Journal of Andrology 2008, 14 (11): 993-7

OBJECTIVE: To construct a short hairpin RNA (shRNA) vector of the hypoxia inducible factor-1alpha (HIF-1alpha), determine its inhibitory effect on the expression of the HIF-1alpha gene in PC-3M cells, and investigate its application prospects in the treatment of prostate cancer.

METHODS: We designed and synthesized the shRNA template sequence specific against HIF-lalpha, inserted it into the vector psilencer 2.1-U6 to generate the plasmid psilencer-HIF, transfected the recombinant plasmid into prostate cancer cell line PC-3M cells and detected the transfection efficiency by cotransfection with the pEGFP vector as well as the expression of HIF-1alpha by RT-PCR and Western blot.

RESULTS: The DNA sequencing analysis showed a complete consistency of the recombinant plasmid psilencer-HIF with the design. Twenty-four hours after the transfection, the rate of transfected plasmid was about (89.26 +/- 4.72)% and the vector-mediated shRNA induced RNA interference (RNAi), while 48 hours transfection reduced the HIF-1alpha mRNA and protein levels by 82.09% and 81.61% respectively (P < 0.01) in PC-3M cells.

CONCLUSION: The shRNA vector was successfully constructed, which can effectively suppress the expression of HIF-1alpha in prostate cancer cells.

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